雷公藤单体T_4体外给药对IL－2R表达的影响

Ｔ_Ⅱ是中草药雷公藤的一类粗提物，临床治疗类风湿关节炎和系统性红斑狼疮等免疫性疾病效果显著。Ｔ_４是进一步从Ｔ_Ⅱ中提取出的单体成分。以正常人外周血单个核细胞（ＰＢＭＣ）为实验材料，测定了不同浓度的Ｔ_Ⅱ（０．０９８～３．１３Ｕ／ｍｌ）和Ｔ_４（０．０３９～５ｎｇ／ｍｌ）对于Ｔ细胞功能的影响。结果表明，Ｔ_Ⅱ和Ｔ_４对Ｔ细胞表面ＩＬ－２Ｒ的表达剂量相关的抑制作用。结果还显示，在药物作用下，ＩＬ－２对ＩＬ－２Ｒ表达的调节作用是很有限的。     

利用噬菌体肽库确定IL-2Rα表位序列①

目的确定IL-2Rα表位,为研制高效特异性强的小分子肽类免疫抑制剂奠定基础。方法用抗IL-2Rα单克隆抗体5G1对噬菌体六肽库进行筛选,选择出与5G1结合较强的克隆,经DNA序列分析,获得保守序列。对含保守序列的克隆进行生物学活性鉴定。结果选择出31个与5G1结合较强的克隆。获得Ser-Ser-Phe和Ser-Ser-Arg两种保守序列。生物学活性鉴定表明含Ser-Ser-Arg序列克隆较含Ser-Ser-Phe被抑制效果好。结论Ser-Ser-Arg是IL-2Rα表位的关键序列。以此表位序列合成的小分子肽可作为IL-2Rα的拮抗剂而成为免疫抑制剂。     

Biochemical and histopathological evaluation of histamine receptors (H1R,H2R,H3R and H4R)-agonist in rabbits

The present study was designed to investigate the biochemical and histopathological changes in the livers of rabbits treated with histamine and histamine receptors (H1R–H4R)-agonist. The cohort comprised of six groups containing five rabbits each. Control group received sterile distilled water (1 mL/kg × b.i.d.) and treated groups received subcutaneous histamine (100 μg/kg × b.i.d.) and H1R–H4R-agonist (histamine trifluoro-methyl toluidide, amthamine, R-[?]-α-methylhistamine, clobenpropit, respectively) each in a dose of 10 μg/kg × b.i.d. (12 h [8 am and 8 pm]) for 30 days. Hepatotoxicity due to these agonists was analyzed using biochemical and histopathological methods. Histamine and H1R–H3R-agonist were found to be hepatotoxic as shown by statistically significant (p < 0.05) elevated levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), most marked in the H3R-agonist group. However, their levels in H4R-agonist group remained very similar to the control group. The entire drug treated groups as compared to control showed significant elevated levels of alkaline phosphatase (ALP). Histopathological examination revealed obvious changes in histamine, H2R- and H3R-agonist groups in terms of alteration of hepatic microstructure, congestion, focal necrosis and increased incidence of multinucleate hepatocytes while H1R and H4R groups showed minimal changes. From the findings of the present study it may be concluded that on repeated administration, histamine and HR-agonists-induced hepatotoxicity which is most pronounced with H3R-agonist though not severe enough to jeopardize the vital functions of liver and warrants further long-term studies.     

IL-4和IL-4受体基因多态性与成人变应性哮喘的关系

目的　研究白细胞介素 4 (IL 4 )、IL 4受体α链的 2个基因多态性位点与中国成人变应性哮喘的关系。方法　采用病例对照方法 ,用聚合酶链反应 限制性片段长度多态性方法 (PCR RFLP)对IL 4启动子区C - 5 89T和IL 4Rα链Q5 76R进行基因分型。结果　IL 4C - 5 89T与中国成人变应性哮喘无关 ,然而 ,变应性哮喘组IL 4Rα链 5 76R R频率显著性高于对照组 (χ2 =9.36 9,P <0 .0 1;OR =3.797) ,且与血浆高IgE相关。结论　IL 4Rα链 5 76R R基因型是中国成人变应性哮喘的基因危险因子     

白细胞介素4和受体基因多态性与儿童变应性哮喘的相关分析

目的：探讨白细胞介素4(IL-4)及受体(IL-4R)基因多态性与儿童变应性哮喘易感性及与血浆总IgE的关系。方法： 采用聚合酶链反应-限制性酶切多态性方法检测IL-4基因启动子区-589位和IL-4R α亚单位Q576R两位点基因多态性，并观察对血浆总IgE的影响。结果：(1)白细胞介素-4基因-589基因多态性与儿童支气管哮喘无相关关系；(2)IL-4受体 α亚单位RR基因型和R 576等位基因频率在儿童支气管哮喘与对照组相比差异显著(P＜0.01，P＜0.01)，且R 576与高IgE相关。结论： 这些数据表明IL-4R α亚单位R 576是中国汉族变应性哮喘患儿的危险因子，且与高IgE相关。     

Leukotriene B4 augments interleukin-2 receptor-beta (IL-2R beta) expression and IL-2R beta-mediated cytotoxic response in human peripheral blood lymphocytes.

Interleukin-2 can induce cytolytic activity in human peripheral blood lymphocytes and this activation is mediated by the beta chain of the interleukin-2 receptor-beta (IL-2R beta). Leukotriene B4 (LTB4) is a potent lipid inflammatory mediator which induces IL-2 and interferon-gamma (IFN-gamma) production from T cells. We examined the ability of LTB4 to modulate IL-2-induced cytolytic activity. Peripheral blood lymphocytes which had been preincubated for 24 hr in the presence of LTB4 responded to 100-fold lower concentrations of IL-2 with an augmentation of natural killer (NK) cell cytotoxic activity. Furthermore, incubation of lymphocytes with graded concentrations of LTB4 augmented the proportion of IL-2R beta+ cells. Peak activity was seen at 10 nM LTB4 and was comparable to that of PHA. By two-colour cytofluorometry, the increased expression of IL-2R beta was found predominantly on CD56+ cells and to a lesser extent on CD8+ cells, while CD4+ cells were unaffected. These observations were correlated at the messenger RNA (mRNA) level with increased IL-2R beta mRNA accumulation following stimulation of purified CD56+ and CD8+ cells with LTB4. CD56-, CD8- cells did not respond to LTB4 by increased IL-2R beta mRNA accumulation. Our data indicate, for the first time, that LTB4 can markedly increase the sensitivity of non-major histocompatibility complex (MHC)-restricted cytotoxic lymphocytes to IL-2, in terms of IL-2-dependent cytotoxic responses, and that this sensitivity is associated with augmented IL-2R beta gene message and cell surface expression.     

IL-2Rbeta links IL-2R signaling with Foxp3 expression

Immunological tolerance to self antigens is a tightly regulated process. Recent work has demonstrated that the forkhead family member Foxp3 is a critical element in the differentiation and function of mouse CD4(+)CD25(+) regulatory T cells (Treg). Recent work has suggested an important role for IL-2 in the development and maintenance of Treg. To directly assess the effect of IL-2 signaling on Treg development and function, we analyzed mice that were genetically deficient in components of the IL-2 receptor (IL-2R). Mice lacking CD25 (IL-2Ralpha) displayed a slight decrease in Treg within the thymus, while peripheral numbers are unchanged. In contrast, we found that mice deficient in CD122 (IL-2Rbeta) had a profound reduction in both thymic and peripheral Treg, coinciding with more rapid development of a fatal lymphoproliferative disease. Expression of a Foxp3 transgene restored Treg and protected against the onset of autoimmunity. Thus, a signal mediated by IL-2Rbeta is essential for the development and homeostasis of Foxp3(+) Treg in vivo.     

Expression of IL-2R, IL-4R, IL-6R on peripheral blood lymphocytes in systemic lupus erythematosus and correlation with disease activity: a prospective study.

AIMS: To study the expression of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R) and interleukin-6 receptor (IL-6R) on peripheral blood lymphocytes (PBL) in patients with systemic lupus erythematosus (SLE); to correlate the level of expression of these receptors with SLE disease activity. METHODS: Peripheral blood lymphocytes were studied by a high sensitivity flow cytometry technique using monoclonal antibodies directed against CD25 (IL-2R alpha chain), CD122 (IL-2R beta chain), CD124 (IL-4R), and CD126 (IL-6R). SLE disease activity was scored using the SLE Disease Activity Index, C3 and C4 concentrations, anti-dsDNA level, and absolute lymphocyte count. RESULTS: Compared with normal controls, PBL from patients with SLE had a higher percentage of CD25+ cells (median 20.8% v 16.5%) and a lower percentage of CD122+ cells (median 13.1% v 22.4%). The difference in CD122+ cells was greater in the CD122weak population than the CD122strong (natural killer cell) population. The percentages of CD124+ and CD126+ PBLs in patients with SLE and controls were similar. On CD25+ cells, the relative antigenic level of the IL-2R alpha chain was significantly higher in patients with SLE (median 2.01 v 1.81). The relative antigenic levels of CD122+, CD124+ and CD126+ cells were similar in patients and controls. Neither the percentages nor the relative antigenic levels of all of these cytokine receptors were correlated with any of the parameters of disease activity. CONCLUSION: Lymphocyte activation in patients with SLE was evident from the increase in CD25 expression on PBL, with a reciprocal decrease in CD122 expression. As the expression of IL-2R, IL-4R, IL-6R did not correlate with disease activity, it seems that these cytokine/receptor systems do not play a direct role in disease activation in SLE.     

老年人淋巴细胞IL-2Rγ链的表达及其意义

目的为探讨IL-2Rγ链在衰老免疫机制中的作用。方法选择了正常青年人8名,健康老年人17名作为研究对象,青年组年龄18～35岁（平均24.9±5.8岁）,老年组年龄65～79岁（平均69.4±4.2岁）,观察了其外周血淋巴细胞对PHA应答增殖程度,CD132、CD45R阳性细胞百分率及时间动力学的变化,同时用自己设计合成的引物采用RT-PCR法对IL-2Rγ链的基因表达进行研究。结果显示老年人淋巴细胞对PHA应答增殖能力低于青年人,分别为0.199±0.055,0.279±0.054,P=0.0012;活化的淋巴细胞表面表达IL-2Rγ链减少,老年人为46.2%±7.7%,青年人为68.0%±11.8%,P<0.001;RT-PCR的结果证实此种改变发生在转录水平;老年人IL-2Rγ链表达的时间动力学曲线与青年人有差异。结论认为IL-2Rγ链对老年人免疫功能衰退可能起了重要的作用。     

Regulatory CD4 T cells: expression of IL-2R alpha chain, resistance to clonal deletion and IL-2 dependency

We recently characterized a CD4+ T cell population expressing the IL-2R alpha chain (CD25), producing IL-10 and resisting clonal deletion induced by viral superantigen (vSAG) encoded by mouse mammary tumor virus [MMTV(SW)]. We now report that these apoptosis-resistant cells are generated in the thymus but not from the immature CD4+ CD8+ thymocytes. They migrate from the thymus and are found in the periphery from at least the 10th day of life, after which they expand with the same kinetics in normal and MMTV(SW)-infected mice. Their strong capacity for expansion in the periphery makes this population insensitive to thymectomy in adulthood. CD4+ CD25+ cells were totally dependent on exogenous IL-2 for growth in vitro and in vivo, and were missing in IL-2 knockout (KO) mice. The absence of this population and/or an inability to produce IL-10 may be the missing link between IL- 2R alpha KO, IL-2 KO and IL-10 KO mice, which all die of inflammatory bowel disease.      

La voie IL-2/IL-2R chez la cellule dendritique module à la fois leur synthèse de cytokines et leur capacité à activer des lymphocytes T CD4 allogéniques

The results provide new insights into the role of IL-2/IL-2R pathway in DC. We report that stimulation of human monocyte-derived DC with LPS strongly upregulated CD25 (α chain of the IL-2R) expression. In addition, by using a humanized monoclonal antibody against CD25, we demonstrated that the IL-2 signalling in DC upregulated both IL-12 and γIFN production but decreased IL-10 synthesis. Anti-CD25 treatment reduced the ability of LPS-DC to induce allogeneic CD4⁺ T cell proliferation as compared to LPS-matured DC. In addition, LPS-matured DC treated with IL-2 had a higher allostimulatory capacity compared to LPS-DC. We also found that LPS-matured DC produced IL-2. Thus, IL-2 seems to contribute actively to DC activation through an autocrine pathway. Moreover, IL-2 pathway in DC is involved in T helper priming. These findings might be useful for protocols in cellular therapy and a valuable tool to understand graft rejection versus the acquisition of peripheral tolerance.     

Tuning inflammation and immunity by the negative regulators IL-1R2 and IL-1R8

Interleukin-1 receptor family members (ILRs) and Toll-Like Receptors (TLRs) are key players in immunity and inflammation and are tightly regulated at different levels. Most cell types, including cells of the innate and adaptive immune system express ILRs and TLRs. In addition, IL-1 family members are emerging as key players in the differentiation and function of innate and adaptive lymphoid cells. IL-1R2 and IL-1R8 (also known as TIR8 or SIGIRR) are members of the ILR family acting as negative regulators of the IL-1 system. IL-1R2 binds IL-1 and the accessory protein IL-1RAcP without activating signaling and can be released as a soluble form (sIL-1R2), thus modulating IL-1 availability for the signaling receptor. IL-1R8 dampens ILR- and TLR-mediated cell activation and it is a component of the receptor recognizing human IL-37. Here, we summarize our current understanding of the structure and function of IL-1R2 and IL-1R8, focusing on their role in different pathological conditions, ranging from infectious and sterile inflammation, to autoimmunity and cancer-related inflammation. We also address the emerging evidence regarding the role of IL-1R8 as a crucial checkpoint molecule in NK cells in anti-cancer and antiviral activity and the potential therapeutic implications of IL-1R8 blockade in specific pathological contexts.     

IL-4R signaling is required to induce IL-10 for the establishment of T(h)2 dominance

The requirement for IL-4 to promote differentiation of naive CD4(+) T cells into T(h)2 effector cell populations was established by classical in vitro studies. More recent in vivo data, however, indicate that signaling through the IL-4R is not essential for acquisition of the T(h)2 phenotype. In order to reconcile these seemingly contradictory conclusions, we have taken advantage of the ability of the excretory/secretory antigens of the gastrointestinal nematode Nippostrongylus brasiliensis to down-regulate T(h)1 cell development and drive T(h)2 cell expansion. We show that the initial development of IL-4-producing T cells is independent of IL-4R signaling but that the subsequent expansion of IL-4-producing CD4(+) T cells in a competitive environment that also contains T(h)1 potential is positively influenced by IL-4R signaling. We find that the production of IL-10 is the key IL-4R-dependent factor required to maintain T(h)2 dominance and that in the absence of IL-4R signaling, T(h)2 expansion can only be achieved by neutralization of T(h)1 cytokines. Moreover, in the absence of IL-4R signaling, reduced IL-10 production is due to the lack in expansion of an IL-10(+) T(h)2 population, rather than a global defect in the production of IL-10 by CD4(+) T cells. Thus, the evolution of T(h)2 dominance is achieved at the expense of T(h)1 cell development, normally restrained by IL-10 in an IL-4R-dependent manner. We conclude that T(h)2 cell development in response to N. brasiliensis antigen requires both IL-4 and IL-10 to act in concert on incipient populations of both T(h)1 and T(h)2 types.     

Immunological,biochemical and histopathological evaluation of histamine receptors (H1R,H2R,H3R and H4R)-antagonist in rabbit experimental model: A short term study

The present study was designed to delineate the immuno- and hepatotoxicological roles of HRs-antagonists in vivo which is elementary in existing literature. The cohort comprised of two experimental studies. Experimental study 1 was designed for immunological investigations and consisted of seven groups and immunized with intravenous injection of SRBC at day 3 containing six rabbits each. Experimental study 2 was designed to assess the functional status of liver and comprised of seven groups containing five rabbits each. In both experimental studies group-I received sterile distilled water intramuscularly, and group II–VI received subcutaneous histamine, pheniramine (H1R-antagonist), ranitidine (H2R-antagonist), iodophenpropit (H3R-antagonist) and JNJ7777120 (H4R-antagonist), respectively while group-VII received DMSO intramuscularly. ELISA was used to assess the immunological investigations. The SRBC-specific immunoglobulins (Igs), IgM and IgG were significantly increased (p < 0.05). Hepatotoxicity due to same histamine and HRs-antagonists were demonstrated by biochemical and histopathological methods. Rabbits in group II–VI had significantly (p < 0.05) elevated levels of serum enzymes (ALT, AST, ALP) and bilirubin. Histopathological examination showed maintained hepatic lobular architecture in histamine and DMSO-treated groups a kin to control. Notable findings in other groups included increased binuclearity in H1R, trinuclearity in H2R, oxyphilic clusters of hepatocytes in H3R and moderate centrilobular necrosis in H3R and H4R-antagonist-treated groups without obvious inflammatory cell infiltration and Kupffer cell prominence. It is concluded that HRs-antagonist play immune suppressive role through H1R, H2R and H4R while immune enhancing role through H3R. In addition, HRs-antagonists appear moderately hepatotoxic in terms of altered serum enzyme levels and non-inflammatory hepatocellular damage.     

Treatment of old mice with IL-2 corrects dysregulated IL-2 and IL-4 production

Splenic T cells from old BALB/c mice, activated in vitro withantibody to CD3e, secrete more IL-4 but less IL-2 than splenicT cells from young mice. The age-associated increase in IL-4secretion is associated with a significantly increased concentrationof intracellular IL-4 and its mRNA, although there is no increasein the number of activated T cells with intracellular IL-4.In contrast, the age-associated decrease in IL-2 secretion isassociated with a significant decrease in the number of activatedT cells with intracellular IL-2. In vivo there is a similarage-associated change in the number of activated T cells withdetectable cytokine. The number of activated T cells with intracellularIL-4 is comparable in old and young mice, while the number ofactivated T cells with intracellular IL-2 is significantly decreasedin old compared with young mice. Of great interest is the factthat old mice continuously exposed to IL-2 In vivo followingthe transplantation of J558 cells expressing the transfectedIL-2 gene product have an increased number of splenic T cellswith intracellular IL-2 that equals the level of such cellsobserved in young mice. Most important, the effect of continuousIL-2 administration in vitro was stable as spleen cells fromold, IL-2-treated mice when stimulated in vitro with anti-CD3ehad a young-like pattern of both intracellular IL-2 and IL-4expression as well as IL-2 and IL-4 secretion following in vitroactivation. Thus, it appears that exposure of old mice to exogenousIL-2 can redress the age-associated imbalance in cytokine expressionin vivo and cytokine secretion in vitro.     

骨肿瘤患者mIL-2R表达和血清sIL-2R水平的观察及临床意义的研究

本实验观察28例恶性骨肿瘤(骨肉瘤14例,软骨肉瘤5例),转移性骨癌4例,,其它5例,及15例良性骨肿患者PBMC 经PHA 刺激后的mIL-2R 表达和血清sIL-2R 水平,设健康对照组15例。实验结果:1.恶性骨肿瘤患者mIL-2R 表达显著降低,在各类型之间则无明显差异;2.恶性骨肿瘤患者sIL-2R水平显著升高,在转移性骨癌明显高于其它类型;3.良性骨肿瘤患者mIL-2R 表达和slL-2R 水平与对照组比较均无明显差异;4.骨肿瘤患者mIL-2R 表达和sIL-2R 水平之间无明确关系。表明mIL-2R 表达不足和sIL-2R 异常升高在恶性骨肿瘤患者机体免疫抑制机制中起重要作用,血清sIL-2R 水平和肿瘤转移及病情变化密切相关;提示sIL-2R 可作为骨肿瘤患者辅助诊断及病情监测的有效指标。